Figure 3: Inhibition of PI3K or Akt abolished the effect of LXR on intracellular protein phosphorylation. MCF7 cells were treated with DMSO and 5 μM GW3965 for 16 h and then cells treated with different inhibitors of PI3K/AKT pathway 30 μM LY-294002 for 30 min, 10 nM Rapamycin for 2 h, 500 nM MK2206 for 2 h, and 1 μM PP242 for 30 min. Cells were harvested to analyze either western immunoblotting by whole cell lysate extraction using various antibodies as described in the figure (A-C) or real time PCR by measuring relative mRNA expression levels for various regulatory gens that control PI3K/AKT pathway (PTEN, PHLPP, and PHLPPL) as described in the figure (D). ABCG1 was used as a key LXR target gen. GAPDH was used as an internal control demonstrating equal protein loading. The data are presented as mean ± SEM. *P < 0.05,***P < 0.001, and #P < 0.0001 versus vehicle by Student's t-test.